Tn5/7-lux: a versatile tool for the identification and capture of promoters in Gram-negative bacteria

Bruckbauer, Steven T; Kvitko, Brian H; Karkhoff-Schweizer, RoxAnn R; Schweizer, Herbert P

doi:10.1186/s12866-015-0354-3

BMC Microbiology

Table 1 Plasmids

From: Tn5/7-lux: a versatile tool for the identification and capture of promoters in Gram-negative bacteria

Plasmid	GenBank accession no.	Pertinent features ^a,b	Source
pTn7xLuxG0	KF532964	Gm^r; luxCDEBA operon transcribed from P _PA4974 contained on DraIII fragment	This Study
pTn7oLuxG0	KF532961	Gm^r; DraIII fragment containing P _PA4974 replaced with DraIII fragment containing P _ompA ^c	This Study
pTn7xLuxG3	KF532965	Km^r; pTn7xLuxG0 with BamHI and PstI sites deleted by partial digestion	This Study
pTn7xLuxK3^d	KC332283	Km^r; pTn7xLuxG3 with Gm^r encoding aacC1 gene replaced with Km^r encoding nptII gene from pFKM4	This Study
pTn7oLuxK3^d	KC332281	Km^r; pTn7xLuxK3 with DraIII fragment containing P _PA4974 replaced with DraIII fragment containing B. pseudomallei P _ompA	This Study
pTn7tLuxK3^d	KC332282	Km^r; pTn7xLuxK3 with DraIII fragment containing P _PA4974 replaced with DraIII fragment containing B. pseudomallei P _tolC	This Study
pTn5/7LuxK3^d	KC332286	Km^r; pTn7xLuxK3 with StuI-DraIII fragment containing P _PA4974 replaced with StuI-DraIII fragment containing Tn5 transposase gene tnpA and flanking mosaic ends	This Study
pTn7oLuxG4	KF532962	Gm^r; pTn7oLuxG0 with BamHI multiple cloning site fragment deleted	This Study
pTn7oLuxK4	KC332284	Km^r; pTn7oLuxG4 with Gm^r encoding aacC1 gene replaced with Km^r encoding nptII gene from pFKM4	This Study
pTn7oLuxT4	KF532963	Tp^r; pTn7oLuxG4 with Gm^r encoding aacC1 gene replaced with Tp^r encoding dhfRII gene from pFTP2	This Study
pTn5/7LuxG4	KF813061	Gm^r; pTn7oLuxG4 with StuI-DraIII fragment containing P _ompA replaced with StuI-DraIII fragment containing Tn5 transposase gene tnpA and flanking mosaic ends	This Study
pTn5/7LuxK4	KF532957	Km^r; pTn7oLuxK4 with StuI-DraIII fragment containing P _ompA replaced with StuI-DraIII fragment containing Tn5 transposase gene tnpA and flanking mosaic ends	This Study
pTn5/7LuxT4	KF813062	Tp^r; pTn7oLuxT4 with with StuI-DraIII fragment containing P _ompA replaced with StuI-DraIII fragment containing Tn5 transposase gene tnpA and flanking mosaic ends	This Study
pTn5/7LuxK5	KF532958	Km^r; pTn5/7LuxK4 with tnpA gene transcribed from P _S12	This Study
pTn5/7LuxT5	KF532959	Tp^r; pTn5/7LuxT4 with tnpA gene transcribed from P _S12	This Study
pTn5/7LuxT6	KC332285	Tp^r; pTn5/7LuxT5 with attB1 inserted at DraIII site	This Study
pTn5/7LuxG6	KC332289	Gm^r; pTn5/7LuxT6 with Tp^r encoding dhfRII gene replaced with Gm^r encoding aacC1 gene from pFGM1	This Study
pTn5/7LuxK6	KC332287	Km^r; pTn5/7LuxT6 with Tp^r encoding dhfRII gene replaced with Km^r encoding nptII gene from pFKM4	This Study
pTn5/7LuxTc6	KC332288	Tc^r; pTn5/7LuxT6 with Tp^r encoding dhfRII gene replaced with Tc^r encoding tetA gene from pFTC2	This Study
pTn7P _DH51-lux	N/A	Km^r; captured DH5α rbsC::Tn5/7LuxK3 insertion on mini-Tn7 delivery plasmid obtained by religation of chromosomal Acc65I fragment	This Study
pTn7P _rbsLuxK4	N/A	Km^r; E. coli rbs promoter (P _rbs) cloned into pTn7LuxK4	This Study
pTn7P _H1ALuxK5	N/A	Km^r; A. baumannii A1S_0945 gene promoter (P _H1A) cloned into pTn7LuxK5	This Study

^aAbbreviations: Ap, ampicillin; FRT, Flp recombinase target; Gm, gentamicin; Km, kanamycin; r, resistance/resistant; Tc, tetracycline; Tp, trimethoprim.
^bSee supplemental methods for details of plasmid constructions.
^cPromoters used are: P _PA4974, P. aeruginosa gene PA4974 promoter; P _ompA, B. pseudomallei ompA promoter; P _tolC, B. pseudomallei tolC promoter; P _S12, B. thailandensis ribosomal S12 gene promoter.
^dPlasmids are missing a FRT site flanking the nptII gene.

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